RU2330034C1 - [4-(5-aminomethyl-2-flourophenyl)-pyperidin-1-yl]-(4-bromine-3-propoxythiophene-2-yl)-methalon hydrochloride as inhibitor of mast cells tryptase - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: described are the bonds of the formula

of pharmaceutical acceptable salt. The bonds possess an inhibiting action in the relation to beta-tryptase. Also, described is the method to treat patients suffering or subject to physiological diseases, with which it is necessary to increase the level of the inhibitor of tryptase.

EFFECT: bonds increase the level of the inhibitor of tryptase.

13 cl, 3 ex, 2 dwg

Description

FIELD OF THE INVENTION

The present invention relates to a substituted arylmethylamine compound, its preparation, a medicament comprising such a compound, its use and its intermediates.

BACKGROUND OF THE INVENTION

Inflammatory processes involving mast cells, in particular asthma, are becoming an increasingly serious public health problem. Asthma is often characterized by the progressive development of a hyperreactive reaction of the trachea and bronchi to both immunospecific allergens and general chemical or physical stimuli, which leads to the formation of chronic inflammation. White blood cells containing IgE receptors, especially mast cells and basophils, are present in the epithelium and the lining smooth muscle tissue of the bronchi. Such leukocytes are initially activated by binding specific respirable antigens to IgE receptors, and then release a number of chemical mediators. For example, mast cell degranulation leads to the release of proteoglycans, peroxidase, arylsulfatase B, chymase and tryptase, which causes narrowing of the bronchioles.

Tryptase is stored in the secretory granules of mast cells and is the main secretory protease of human mast cells. Tryptase is involved in a variety of biological processes, including the degradation of vasodilating and bronchodilating neuropeptides (Caughey et al., J. Pharmacol. Exp. Ther., 1988, 244, pp. 133-137; Franconi et al., J. Pharmacol. Exp . Ther., 1988, 248, pp. 947-951; Tam et al., Am. J. Respir. Cell Mol. Biol., 1990, 3, pp. 27-32) and modulation of the bronchial response to histamine (Sekizawa et al., J. Clin. Invest., 1989, 83, pp. 175-179).

Therefore, tryptase inhibitors may be useful as anti-inflammatory agents (K Rice, PA Sprengler, Current Opinion in Drug Discovery and Development, 1999, 2 (5), pp. 463-474), especially in the treatment of chronic asthma (MQ Zhang, H. Timmerman, Mediators Inflamm., 1997, 112, pp. 311-317), and may also be useful in the treatment or prevention of allergic rhinitis (SJ Wilson et al., Clin. Exp. Allergy, 1998, 28, pp. 220-227 ), chronic intestinal irritability syndrome (SC Bischoff et al., Histopathology, 1996, 28, p. 1-13), psoriasis (A. Naukkarinen et al., Arch. Dermatol. Res., 1993, 285, p. 341- 346), conjunctivitis (AAIrani et al., J. Allergy Clin. Immunol., 1990, 86, p. 34-40), atopic dermatitis (A. Jarvikallio et al., Br. J. Dermatol., 1997, 136, pp. 871-877), rheumatoid arthritis (LC Tetlow et al., Ann. Rheum. Dis., 1998, 54, p. 549 -555), osteoarthritis (MG Buckley et al., J. Pathol., 1998, 186, pp. 67-74), gouty arthritis, rheumatoid spondylitis and diseases of the destruction of articular cartilage.

In addition, tryptase has been shown to be an effective mitogen for fibroblasts, suggesting its involvement in pulmonary fibrosis in asthma and interstitial pulmonary processes (Ruoss et al., J. Clin. Invest., 1991, 88, pp. 493-499 )

Therefore, tryptase inhibitors may be useful in the treatment or prevention of fibrotic diseases (JA Cairns and AF Walls, J. Clin. Invest., 1997, 99, pp. 1313-1321), for example, fibrosis, scleroderma, pulmonary fibrosis, liver cirrhosis, myocardial fibrosis , neurofibroma and hypertrophic scars.

In addition, tryptase inhibitors may be useful in the treatment or prevention of myocardial infarction, stroke, angina pectoris and other consequences of rupture of atherosclerotic plaques (M. Jeziorska et al., J. Pathol., 1997, 182, pp. 115-122).

It was also found that tryptase activates prostromelysin, which, in turn, activates collagenase, thereby initiating the destruction of cartilage and periodontal connective tissue, respectively.

Therefore, tryptase inhibitors may be useful in the treatment or prevention of arthritis, periodontal disease, diabetic retinopathy and tumor growth (W.J. Beil et al., Exp. Hematol., (1998) 26, p. 158-169). Tryptase inhibitors may also be useful in the treatment of anaphylaxis (LB Schwarz et al., J. Clin. Invest., 1995, 96, pp. 2702-2710), multiple sclerosis (M. Steinhoff et al., Nat. Med. (NY ), 2000, 6 (2), pp. 151-158), peptic ulcers and syncytial viral infections.

Substituted arylmethylamines represented by compounds of formula (A), their preparation, pharmacological preparations containing such compounds, and their pharmaceutical use in the treatment of painful conditions that can be corrected by inhibition of tryptase are set forth in pending US patent application No. 09/843126. The general information about compounds of formula (A) in US patent application No. 09/843126 also includes a compound of the present invention of formula I. However, US patent application No. 09/843126 does not specifically disclose information about a compound of formula I.

Accordingly, there is a need for a new and useful compound that has particularly valuable pharmaceutical properties in its ability to inhibit tryptase. Such a compound should easily find use in the treatment of patients suffering from diseases that can be alleviated by the administration of a tryptase inhibitor, for example, inflammatory conditions involving mast cells, inflammation and diseases or disorders associated with the degradation of vasodilating and bronchorelaxing neuropeptides.

SUMMARY OF THE INVENTION

The present invention relates to a compound of formula I

or a prodrug, a pharmaceutically acceptable salt or solvate of the compound.

In addition, the present invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula I and a pharmaceutically acceptable carrier.

In addition, the present invention relates to the use of a compound of formula I as a tryptase inhibitor, comprising administering the compound in a composition comprising tryptase. The present invention also relates to the use of a compound of formula I for the treatment of patients suffering from or susceptible to physiological diseases in which an increase in the level of tryptase inhibitor is required, the treatment comprising administering to the patient a therapeutically effective amount of the compound according to paragraph 1.

The present invention also relates to a method for producing a compound of formula I.

Brief Description of the Drawings

Aspects, features and advantages of the present invention will be more apparent from the following detailed descriptions, taken in conjunction with the accompanying drawings, which are provided for illustration only and are not restrictive to the present invention, these include the following.

Figure 1: levels of compound I in plasma, bronchoalveolar lavage (BAL) and lungs, measured 2 hours after administration of compound I at a dose of 1 mg / kg orally. Values are mean ± standard error for 6-8 animals.

Figure 2: levels of compound I in plasma and lungs, measured 24 hours after administration. Values are mean ± standard error of 3-4 animals.

FULL DESCRIPTION OF THE INVENTION

Definitions

The terms used above and throughout the text of the present description and the attached claims, unless otherwise indicated, have the following accepted meanings.

As used herein, the term “compound of the present invention” and equivalent expressions encompass a compound of formula I as described above and includes a prodrug, a pharmaceutically acceptable salt and a solvate, for example a hydrate. Similarly, reference to intermediates, whether included in the claims themselves, encompasses salts and solvates in cases where context permits. For complete clarity, specific cases where this is permitted by the context are sometimes indicated in the text, but such cases are purely illustrative and do not involve the exclusion of other cases where the context allows such an interpretation.

The term “treating” or “treating” as used herein includes prophylactic therapy as well as treating a diagnosed condition of a patient.

“Patient” means a human or other mammal.

An “effective amount” is intended to describe an amount of a compound effective in creating the desired therapeutic effect.

“Prodrug” means a compound that is suitable for administration to a patient, does not have undesirable toxicity, does not cause irritation, an allergic reaction, and the like. and during metabolic processes in the body (for example, hydrolysis) can be converted into a compound of the present invention. A detailed discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A. C. S. Symposium Series, as well as Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.

Specific or preferred embodiments

In addition, the present invention relates to the use of a compound of formula I for the treatment of patients suffering from physiological diseases that can be alleviated by administering to a patient a therapeutically effective amount of a compound of formula I. Specific embodiments for physiological diseases that can be treated with a compound of the present invention include, but Of course, not limited to, inflammatory diseases such as joint inflammation, arthritis, rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, psoriatic arthritis and other chronic inflammatory joint diseases. Other embodiments for physiological diseases that can be treated using the present invention include physiological diseases such as chronic obstructive pulmonary disease (COPD), exacerbation of COPD, destruction of articular cartilage, ocular conjunctivitis, spring conjunctivitis, irritable bowel syndrome, asthma, allergic rhinitis, interstitial pulmonary processes, fibrosis, scleroderma, pulmonary fibrosis, cirrhosis, myocardial fibrosis, neurofibromas, hypertrophic scars, various dermatological diseases such as atopic dermatitis and psoriasis, myocardial infarction, stroke, angina pectoris and other consequences of rupture of atherosclerotic plaques, as well as periodontal disease, diabetic retinopathy, tumor growth, anaphylaxis, multiple sclerosis, peptic ulcers and syncytial viral infections.

In one specific embodiment, the present invention relates to the use of a compound of formula I for the treatment of patients with asthma, which comprises administering to the patient a physiologically effective amount of a substance.

In another specific embodiment, the present invention relates to the use of a compound of formula I for the treatment of patients suffering from COPD, which comprises administering to the patient a physiologically effective amount of a substance.

In another specific embodiment, the present invention relates to the use of a compound of formula I for the treatment of patients suffering from an exacerbation of COPD, which comprises administering to the patient a physiologically effective amount of a substance.

In another specific embodiment, the present invention relates to the use of a compound of formula I for the treatment of patients suffering from allergic rhinitis, which comprises administering to the patient a physiologically effective amount of a substance.

In another specific embodiment, the present invention relates to the use of a compound of formula I for the treatment of patients suffering from joint inflammation, which comprises administering to the patient a physiologically effective amount of a substance.

In another specific embodiment, the present invention relates to the use of a compound of formula I for the treatment of patients suffering from irritable bowel syndrome, which comprises administering to the patient a physiologically effective amount of a substance.

In addition, the present invention extends to a pharmaceutical composition comprising a compound of formula I, a second compound selected from the group consisting of a beta adrenergic agonist, anticholinergic agent, anti-inflammatory corticosteroid and anti-inflammatory agent, and a pharmaceutically acceptable carrier thereof. In such a composition, the compound of formula I and the second compound are present in amounts that provide a therapeutically effective activity, i.e., an additive or synergistic effect. Specific inflammatory diseases or disorders that can be treated with such a pharmaceutical composition, among others, include asthma.

In addition, the present invention relates to a method of treating patients suffering from inflammatory disorders, which comprises administering to a patient a compound of formula I and a second compound selected from the group consisting of a beta adrenergic agonist, anticholinergic agent, anti-inflammatory corticosteroid and anti-inflammatory agent. In such a method, the compound of formula I and the second compound are present in amounts that provide a therapeutically effective activity, that is, an additive or synergistic effect. In such a method of the present invention, the compound of the present invention can be administered to the patient before the second compound, and the second compound can be administered to the patient before the connection of the present invention or the compound of the present invention and the second compound can be administered simultaneously. Specific examples of adrenergic agonists, anticholinergics, anti-inflammatory corticosteroids and anti-inflammatory agents used in accordance with this method are described below.

Pharmacological preparations

As noted above, the compound of the present invention exhibits useful pharmacological activity and, accordingly, can be included in pharmaceutical compositions and used in the treatment of patients suffering from certain medical disorders. Thus, in accordance with a further aspect, the present invention provides pharmaceutical compositions comprising a compound of the invention and a pharmaceutically acceptable carrier thereof. The term "pharmaceutically acceptable", as used herein, preferably means approved by the legislative authority of a government, in particular the federal or state government, or introduced into the US Pharmacopeia or other generally accepted pharmacopeia for use in animals and, in particular, in humans. Suitable pharmaceutical carriers are described in E.W. Martin "Remington's Pharmaceutical Sciences".

In accordance with the present invention, the pharmaceutical compositions can be prepared according to conventional methods using one or more pharmaceutically acceptable excipients or excipients. Excipients include, but are not limited to, diluents, fillers, binders, disintegrants, glidants, lubricants, surfactants, sterile aqueous media, and various non-toxic organic solvents. The compositions may take the form of tablets, capsules, pills, sustained release formulations, granules, powders, aqueous solutions or suspensions, injection solutions, elixirs or syrups, and may contain one or more agents selected from the group consisting of sweeteners, perfumes, colorants or stabilizers in order to receive pharmaceutically acceptable preparations. The choice of carrier and the content of active substance in the carrier is usually determined in accordance with the solubility and chemical properties of the active compound, the particular route of administration and the provisions that must be observed in pharmaceutical practice. For example, excipients such as lactose, microcrystalline cellulose, gelatinized starch, unmodified starch, silicified microcrystalline cellulose, mannitol, sorbitol, xylitol, dextrates, fructose, sodium citrate, calcium carbonate, dicalcium phosphate dihydrate, calcium sulfate along with binders such as polyvinylpyrrolidone, hydroxypropyl methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, methyl cellulose, sodium salt arboxymethyl cellulose, gelatinized starch, starch, polyethylene glycols, polyethylene oxide, polycarbophiles, gelatin and gum, and disintegrating agents such as croscarmellose sodium, starch glycolate sodium, crospovidone, starch-silica, starch such as magnesium stearate, calcium stearate, stearic acid, hydrogenated vegetable oil, mineral oil, polyethylene glycols, glycerol esters and fatty acids x acids, sodium lauryl sulfate and glidants, such as silicon dioxide, talc, starch, along with certain surfactants such as sodium lauryl sulfate, sorbitan esters, polyoxyethylene fatty acid esters, poloxamer, polyoxyethylene ether, sodium docusate, polyethoxylated castor oil and benzalkonium chloride. For the preparation of the capsule, it is convenient to use fillers such as lactose, microcrystalline cellulose, pregelatinized starch, unmodified starch, pure silicified microcrystalline cellulose or a mixture of two or more fillers with or without the addition of binders listed above, along with a suitable surface-active agent (agents), disintegrants, facilitating the sliding of substances, lubricants, etc., as indicated above. When using aqueous suspensions, they may contain emulsifiers or substances that contribute to the formation of suspensions. Diluents may also be used, for example, sucrose, ethanol, polyethylene glycol, propylene glycol, glycerin and chloroform, or mixtures thereof. Such pharmaceutically acceptable carriers may also be sterilized water and oils, including those of petroleum, animal, vegetable or synthetic origin, for example peanut oil, soybean oil, mineral oil, sesame oil and the like. When administered intravenously, the preferred carrier is water. Saline solutions and aqueous solutions of dextrose and glycerol can also be used as liquid carriers, especially for injectable solutions. Suitable pharmaceutical excipients include mannitol, human serum albumin, starch, glucose, lactose, gelatin, malt, rice, flour, chalk, silica gel, magnesium carbonate, magnesium stearate, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerin , propylene, glycol, water, ethanol and the like. Such formulations may take the form of solutions, suspensions, tablets, pills, capsules, powders, sustained release dosage forms, and the like.

Naturally, the pharmaceutical composition of the present invention will contain a therapeutically effective amount of the active substance along with an appropriate amount of carrier so as to provide a form for proper administration to the patient. Although intravenous injection is a very effective form of administration, other methods can be used, for example, injection or oral, nasal or parenteral administration, which are discussed below.

Treatment methods

The compound of formula I has the ability to inhibit tryptase in accordance with the tests described in the literature and described below in the present description, and the results of such tests are presumably correlated with pharmacological activity in humans and other mammals. Thus, in yet another embodiment, the present invention relates to the use of a compound of formula I or a composition containing it for the treatment of patients suffering or prone to painful conditions that can be alleviated by the administration of a tryptase inhibitor. For example, a compound of formula I is useful for treating inflammatory diseases such as joint inflammation, including arthritis, rheumatoid arthritis, and other types of arthritis, such as rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, psoriatic arthritis, osteoarthritis, or other chronic inflammatory joint disease or joint cartilage destruction disease, ocular conjunctivitis, spring conjunctivitis, irritable bowel syndrome, asthma, allergic rhinitis, interstitial e pulmonary processes, fibrosis, scleroderma, pulmonary fibrosis, liver cirrhosis, myocardial fibrosis, neurofibromas, hypertrophic scars, various dermatological diseases, such as atopic dermatitis and psoriasis, myocardial infarction, stroke, angina pectoris and other consequences of rupture of atherosclerotic plaques, as well as period diabetic retinopathy, tumor growth, anaphylaxis, multiple sclerosis, peptic ulcers or syncytial viral infections.

According to other embodiments of the invention, it provides a method of treating a patient, human or animal suffering or susceptible to diseases that can be alleviated by the administration of a tryptase inhibitor, for example, the diseases described above, and such treatment comprises administering to the patient an effective amount of a compound of the invention or a preparation containing a compound of the invention.

Combination therapy

As explained above, other pharmaceutically active substances may be used in combination with the compound of formula I, depending on the disease being treated. For example, in the treatment of asthma, beta-adrenergic agonists, such as albuterol, terbutaline, formoterol, fenoterol or prenaline, as well as anticholinergics, such as ipratropium bromide, anti-inflammatory corticosteroids, such as beclomethasone dipropionone diacetone dimethyl acetone tricinol, may be included, agents such as sodium cromoglycate and nedocromil sodium. Thus, the present invention extends to a pharmaceutical composition comprising a compound of formula I and a second compound selected from the group consisting of a beta adrenergic agonist, anticholinergic agent, anti-inflammatory corticosteroid and anti-inflammatory agent, and a pharmaceutically acceptable carrier thereof. Specific pharmaceutical carriers useful for a given pharmaceutical composition are described herein.

In addition, the present invention relates to a method for treating patients suffering from asthma, which comprises administering to a patient a compound of the present invention and a second compound selected from the group consisting of a beta adrenergic agonist, anticholinergic agent, anti-inflammatory corticosteroid and anti-inflammatory agent. In such a combined method, the compound of the present invention may be administered before the second compound, the compound of the present invention may be administered after the second compound or the compound of the present invention and the second compound may be administered simultaneously.

Delivery Methods

In accordance with the present invention, a compound of formula I or a pharmaceutical composition comprising this compound can be administered parenterally, through the mucous membrane, for example, orally, nasally, pulmonally or rectally, or in a transdermal manner.

Oral delivery

The present description provides oral solid dosage forms for use, a general description of which is given in Remington's Pharmaceutical Sciences, 18th Ed. 1990 (Mack Publishing Co. Easton PA 18042), chapter 89, which is incorporated herein by reference. Solid dosage forms include tablets, capsules, pills, lozenges or lozenges, cachets or pellets. For the preparation of such preparations can be used liposomal or proteinoid encapsulation (such as, for example, proteinoid microspheres, presented in patent U.S. Patent No. 4925673). Liposome encapsulation can be used, and derivatives of liposomes and various polymers can be obtained (e.g., U.S. Patent No. 5013556). Possible solid dosage forms for therapeutic use are described in Marshall, K. In: Modern Pharmaceutics Edited by G.S. Banker and C.T. Rhodes, chapter 10, 1979, which is incorporated herein by reference. In the General case, the composition will include a compound of the present invention and inert ingredients that provide protection from the environment of the stomach and the release of biologically active material, that is, the compounds of the present invention, in the intestine.

In addition, oral dosage forms of a compound of the present invention are specifically provided. Such a compound may be chemically modified so that oral delivery is more effective. In general, the proposed chemical modification involves the attachment of at least one functional fragment to the component molecule itself, which functional fragment provides (a) inhibition of proteolysis and (b) absorption into the bloodstream from the stomach or intestines. In addition, it is desirable to increase the overall stability of the compounds of the present invention and increase the circulation time in the body. Examples of such functional fragments include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone and polyproline. Abuchowski and Davis, 1981, "Soluble Polymer-Enzyme Adducts" In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367-383; Newmark et al., 1982, J. Appl. Biochem. 4: 185-189. Other polymers that can be used include poly-1,3-dioxolane and poly-1,3,6-trioxocane. As noted above, functional fragments containing polyethylene glycol are preferred for pharmaceutical use.

For the compound of the present invention, the release site may be the stomach, small intestine (duodenum, jejunum or ileum) or large intestine. Those skilled in the art will recognize available formulations that will not dissolve in the stomach, but will release the substance in the duodenum or in another area of the intestine. Preferably, the release will avoid the negative effects of the stomach environment either by protecting the compound of the present invention or by releasing the substance outside the stomach environment, for example, in the intestine.

To ensure absolute stability in the stomach, a coating is impervious to at least pH 5.0. Examples of the most common inert ingredients that are used as enteric coatings are acetyl cellulose trimellitate (TAC), hydroxypropyl methyl cellulose phthalate (GPMCF), GPMCF 50, GPMCF 55, polyvinyl acetate phthalate (PVAF), Eudragit L30D, Efalicyl acetate, Aquatic Phalate Acetate (Aquatic), Aquater Phalate Acetate (Aquatic), Aquater L, Eudragit S and shellac. Such coatings may be used in the form of mixed films.

Coatings or a mixture of coatings can also be used on tablets that are not intended to protect against the environment of the stomach. These may include sugar coatings or coatings that make swallowing tablets easier. Capsules may consist of a hard shell (e.g. gelatin) for delivery of a dry therapeutic agent, i.e. powder; for liquid forms, a soft gelatin shell may be used. The wrapper material for the wafers may be thick starch or other edible paper. Moisture concentration techniques can be used to prepare pills, lozenges, molded tablets, or tablet powders.

A therapeutic agent may be included in the form of aggregates of small particles in the form of granules or pellets with a particle size of about 1 mm. The composition of the capsule administration material may also be a powder, lightly pressed tabs, or even tablets. A therapeutic agent can be prepared by compression.

Dyes and perfumes may be included. For example, a dosage form of a compound of the present invention (for example, encapsulation in liposomes or microspheres) can be prepared and then introduced into a food product, such as a chilled beverage containing colorants and perfumes.

You can dilute or increase the volume of the therapeutic agent due to inert material. Such diluents may include carbohydrates, especially mannitol, alpha-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch. Some inorganic salts can also be used as fillers, including calcium triphosphate, magnesium carbonate and sodium chloride. A range of commercially available diluents include Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell.

Disintegrants may be included in a therapeutic agent in the form of a solid dosage form. Materials used as disintegrants include, but are not limited to, starch based industrial disintegrants, such as Explotab. Starch glycolate sodium, amberlite, carboxymethyl cellulose sodium, ultramilopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may be used. Another form of disintegrants is insoluble cation exchange resins. Powdered gums can be used as disintegrants and binders, and these may include powdered gums such as agar, karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

Binders can be used to form a therapeutic agent in a hard tablet, and such binders include materials from natural products, such as gum, tragacanth, starch and gelatin. Other binders include methyl cellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). To granulate the therapeutic agent, polyvinylpyrrolidone (PVP) and hydroxypropyl methylcellulose (HPMC) in alcohol solutions can be used.

A friction-reducing agent may be included in the composition of the therapeutic agent in order to prevent sticking during the preparation of the composition. Lubricants can be used as a layer between the therapeutic agent and the matrix wall, and, among others, may include stearic acid, including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000 may also be used.

Glidants may be added that improve the fluidity of the drug in the preparation of the composition and facilitate reorganization during molding. Glidants include starch, talc, pyrogenic silica and hydrated aluminosilicate.

To aid dissolution of the therapeutic agent in the aquatic environment, a surfactant may be added as a wetting agent. Surfactants may include anionic detergents, for example sodium lauryl sulfate, sodium dioctyl sulfosuccinate and sodium dioctyl sulfonate. It is possible to use cationic detergents, which may include benzalkonium chloride and benzetomium chloride. The list of possible non-ionic detergents that may be included as surfactants in the preparation includes lauromacrogol 400, polyoxyl stearate 40, polyoxyethylene derivative of hydrogenated castor oil 10, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, fatty sugar ether acids, methyl cellulose and carboxymethyl cellulose. Such surfactants may be present in the preparation of a compound of the present invention both in pure form or in a mixture in various ratios.

Supplements that are able to enhance the absorption of the compounds of the present invention include, for example, fatty acids: oleic acid, linoleic acid and linolenic acid. A controlled-release oral preparation may be desirable. The drug may be incorporated into an inert matrix that provides release either by diffusion or by leaching mechanisms, such as gum. The composition of the drug may also include slowly decaying matrices. A number of enteric coatings also have a delayed release effect.

Another form of controlled release of this therapeutic agent is a method based on the OROS therapeutic system (Alza Corp.), that is, the drug consists of a semipermeable membrane that allows water to penetrate and, due to osmotic effects, expels the drug through a single thin lumen.

Other coatings may be used to prepare the preparation. These include a variety of sugars that can be applied in a coating bath. The therapeutic drug can also be administered in a film-coated tablet, and the materials used for this are divided into 2 groups. The first is non-enteric materials, and it includes methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, methyl hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose sodium, providone and polyethylene glycols. The second group consists of enteric materials, which are usually represented by phthalic acid esters.

A mixture of materials may be used to provide an optimal film coating. The film coating may be applied in a coating bath or in a fluidized bed, or by compression coating.

Pulmonary Delivery

The present document also provides pulmonary delivery of a compound of the present invention, either individually or in the form of a pharmaceutical composition. The compound is delivered to the lungs of mammals by inhalation and is transported through the lining pulmonary epithelium into the bloodstream. Other reports of this include Adjei et al., 1990, Pharmaceutical Research, 7: 565-569; Adjei et al., 1990, International Journal of Pharmaceutics, 63: 135-144 (leuprolide acetate); Braquet et al., 1989, Journal of Cardiovascular Pharmacology, 13 (suppl. 5): 143-146 (endothelin-1); Hubbard et al., 1989, Annals of Internal Medicine, Vol. III, pp. 206-212 (a1-antitrypsin); Smith et al., 1989, J. Clin. Invest. 84: 1145-1146 (a-1 proteinase); Oswein et al., 1990, "Aerosolization of Proteins", Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March (recombinant human growth hormone); Debs et al., 1988, J. Immunol. 140: 3482-3488 (interferon-gamma and tumor necrosis factor alpha) and Platz et al., U.S. Patent No. 5,284,656 (colony stimulating factor of granulocytes). The method and composition for the pulmonary delivery of systemic drugs is described in U.S. Patent. Patent No. 5451569, issued September 19, 1995 by Wong et al.

For practical use, the present invention provides a wide range of mechanical devices for the pulmonary delivery of therapeutic drugs, including, among others, aerosol devices, metered-dose inhalers and powder inhalers, all of which are known to those skilled in the art.

Some specific examples of commercially available devices suitable for the practice of the present invention include an Ultravent aerosol apparatus manufactured by Mallinckrodt, Inc., St. Louis, Missouri; Acorn II aerosol apparatus manufactured by Marquest Medical Products, Englewood, Colorado; Ventolin metered-dose inhaler manufactured by Glaxo Inc., Research Triangle Park, North Carolina, and Spinhaler powder inhaler manufactured by Fisons Corp., Bedford, Massachusetts, and many others.

All such devices require the use of preparations suitable for dispensing a compound of the present invention. Each preparation is generally suitable for use with the particular type of device used, and may include the use of an appropriate propellant in addition to conventional diluents, excipients and / or carriers suitable for the treatment. It is also proposed to use liposomes, microcapsules or microspheres, inclusion complexes or other types of carriers. The chemically modified compound of the present invention may also be formulated in various preparations depending on the type of chemical modification or the type of device used.

Formulations suitable for use in an aerosol apparatus, both in a jet and in an ultrasound apparatus, will usually comprise a compound of the present invention dissolved in water at a concentration of about 0.1 to 25 mg of the compound per 1 ml of solution. The drug may also include a buffer and simple sugar (for example, to stabilize and regulate the osmotic pressure). The composition in the aerosol apparatus may also contain a surfactant to reduce or prevent surface-stimulated aggregation of the compound caused by spraying the solution during aerosol formation.

Formulations of a preparation for use in a metered-dose inhalation device will typically contain finely divided powder containing a compound of the invention suspended in a propellant with a surfactant. The propellant can be any conventional material that is used for these purposes, for example, chlorofluorocarbon, hydrochlorofluorocarbon, hydrofluorocarbon or hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol and 1,1,1,2-tetrafluoroethane or combinations thereof. Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid may also be suitable as a surfactant.

Dosage formulations using a powder inhalation device will contain finely divided powder containing a compound of the present invention, and may also include an excipient, for example, lactose, sorbitol, sucrose or mannitol, in amounts that facilitate spraying of the powder from the device, for example, from 50 to 90 wt.% the drug. The compound of the present invention should advantageously be prepared in ground form with an average particle size of less than 10 mm (or microns), preferably between 0.5 and 5 mm, for the most efficient delivery to the peripheral lungs.

Nasal delivery

Nasal delivery of a compound of the present invention is also provided. Nasal delivery ensures that the compound enters the bloodstream immediately after administration of the therapeutic drug into the nose without the need to deposit the product in the lungs. Preparations for nasal delivery include formulations with dextran or cyclodextran.

Transdermal delivery

The present invention finds application in a wide variety of methods for transdermal administration of a drug, for example, through a transdermal patch. Transdermal patches are described, for example, in U.S. Patent No. 5407713, issued April 18, 1995 by Rolando et al .; U.S. Patent No. 5352456, issued October 4, 1994, Fallon et al .; U.S. Patent No. 5332213, issued August 9, 1994 by D'Angelo et al .; U.S. Patent No. 5336168, issued August 9, 1994 to Sibalis; U.S. Patent No. 5290561, issued March 1, 1994, Farhadieh et al .; U.S. Patent No. 5254346, issued October 19, 1993 by Tucker et al .; U.S. Patent No. 5164189, issued November 17, 1992, Berger et al .; U.S. Patent No. 5163899, issued November 17, 1992 to Sibalis; U.S. Patent Nos. 5088977 and 5087240, issued February 18, 1992 to Sibalis; U.S. Patent No. 5008110, issued April 16, 1991 to Benecke et al., And U.S. Patent No. 4921475, issued May 1, 1990 by Sibalis, the disclosure of information for each of which, by reference to them, is fully incorporated in this document.

It can easily be established that the transdermal route of administration can be made more effective by the use of an agent that enhances penetration through the skin, for example, agents described in U.S. Patent No. 5164189 (above), U.S. Patent No. 5008110 (above) and U.S. Patent No. 4879119, issued November 7, 1989 by Aruga et al., The disclosure of information for each of which, by reference to them, is fully included in this document.

Topical application

For topical use, gels (water or alcohol based), creams or ointments containing the compounds of the invention can be used. The compounds of the invention may also be formulated in a gel or matrix base for use in a patch, which will provide controlled release of the compound through the transdermal barrier.

Rectal introduction

Solid compositions for rectal administration include suppositories formulated according to known methods and containing a compound of the invention.

Dosage

The percentage of active component in the compositions of the invention may vary, it being necessary that it contains such a fraction that would provide an appropriate dosage. It is understood that several unit dosage forms can be administered at about the same time. The dose used will be determined by the doctor and depends on the desired therapeutic effect, method of administration and duration of treatment, as well as the patient's condition. For adults, the doses are usually of the order of 0.001-50, preferably of the order of 0.001-5 mg / kg of body weight when administered by inhalation, of the order of 0.01-100, preferably of 0.1 to 70, more specifically of 0.5 to 10 mg / kg body weight per day for oral use and on the order of 0.001-10, preferably from 0.01 to 1 mg / kg body weight per day for intravenous administration. In each case, the dose will be determined in accordance with the factors specific to the treatment, for example age, weight, general health and other features that may affect the effectiveness of the drug.

In addition, in accordance with the invention, the compound can be administered as often as necessary to achieve the desired therapeutic effect. Some patients may show a quick response to a higher or lower dose, and much lower doses may be sufficient for them. For other patients, long-term courses of treatment with a frequency of 1 to 4 doses per day may be necessary in accordance with the physiological needs of each individual patient. In general, the active substance can be administered orally from 1 to 4 times a day. Of course, for some patients it will be necessary to establish no more than one or two doses per day.

Naturally, a patient for whom administration of a compound of the present invention is an effective therapeutic regimen is preferably a human, but can be any animal. Thus, as can be easily ascertained by any person skilled in the art, the methods and pharmaceutical compositions of the present invention are particularly suitable for administration to any animal, in particular a mammal, including, but not limited to, domestic animals, for example, belonging to the feline or canine family agricultural animals, for example, among others, cows, horses, goats, sheep and pigs, wild animals (in the wild or in a zoological garden), experimental animals, for example mice, rats, rabbits, goats, sheep, pigs, dogs, cats, etc., birds, such as chickens, turkeys, songbirds, that is, for use in veterinary medicine.

Cooking Details

The compound of formula I can be obtained by applying or adapting known methods, which means methods that have been previously used or described in the literature, for example, described in R.C. Larock in Comprehensive Organic Transformations, VCH publishers, 1989, or as described herein.

In the reactions described below, it may be necessary to protect the reaction functional groups, for example amino groups, in order to avoid their undesirable participation in the reactions. Conventional protecting groups may be used in accordance with accepted practice, for examples, see T.W. Greene and P.G.M. Wuts in "Protective Groups in Organic Chemistry" John Wiley and Sons, 1991.

In particular, a compound of formula I can be obtained as shown in schemes 1-3.

For example, the compound of the present invention is an achiral compound, the preparation of which involves convergent synthesis. Scheme 1 below shows the methods whose final step is the preparation of amine 10. The following Scheme 2 contains methods that produce acid 16. Scheme 3 below shows the methods that produce a compound of formula I, and the synthesis of a compound of formula I involves a two-step process. sequence. Getting 10, 16 and the compounds of formula I of the present invention are alternately discussed below.

Compound 2 is converted to compound 3 by protecting the amino group with an amine protecting agent, for example, 1,2-bis (chlorodimethylsilyl) ethane, in the presence of a tertiary amine, for example triethylamine, in a suitable inert solvent, for example dichloromethane, to give the protected compound 3.

Compound 3 is converted to compound 5 by alkylation of compound 4 using compound 3 under conditions of alkylation reactions involving a strong base, for example n-butyllithium, in a suitable aprotic solvent, for example tetrahydrofuran, so as to obtain a hydroxyl derivative, compound 5.

Compound 5 is converted to compound 6 by deprotecting its amino group with a deprotecting agent, for example, a strong inorganic acid, such as phosphoric acid, in the presence of an inert solvent, for example heptane, to give compound 6 without protection.

Compound 6 is converted to compound 7 by dehydration using a strong inorganic acid, such as phosphoric acid, and subsequent neutralization of the product with a strong inorganic base, such as aqueous sodium hydroxide, to give dehydrated compound 7.

Compound 7 is converted to compound 8 by introducing the protection of an amino group with an amine protecting agent, for example BOC anhydride, in a mixed aqueous-organic solvent system in which the organic solvent is a polar organic solvent, for example methanol, using a strong inorganic base, for example aqueous sodium hydroxide so as to obtain a protected BOC compound 8.

Compound 8 is converted to compound 9 by hydrogenation with a reducing agent, for example palladium hydroxide on a carbon substrate (20%), in a mixed solvent system, for example methanol acetic acid, to give a deprotected piperidine salt, compound 9.

Compound 9 is converted to its free base form, compound 10, by neutralization using a strong inorganic base, for example aqueous sodium hydroxide, so as to obtain the final compound 10.

Carbon disulphide is converted to compound 11 by acylation using an acylating agent, propyl chloroformate, in the presence of a strong inorganic base, for example potassium hydroxide, in n-propanol, so as to obtain compound 11.

Compound 11 is converted to compound 12 by alkylation of acetone with compound 11 in the presence of a strong base, for example sodium hydride, to give an alkylated compound 12.

Compound 12 is converted to compound 13 by alkylating compound 12 with alpha bromomethyl acetate in the presence of a tertiary amine, for example triethylamine, to give an alkylated compound 13.

Compound 13 is converted to compound 14 by cyclization in a strongly basic medium, for example sodium methoxide, in the presence of a protic organic solvent, for example methanol, to give cyclic compound 14.

Compound 14 is converted to compound 15 by bromination in an inert organic solvent, for example trichloromethane or in a mixture of tert-butyl methyl ether / heptane, to give brominated compound 15.

Compound 15 is converted to compound 16 by hydrolysis using a strong inorganic base, such as lithium hydroxide.

Compound 16 is converted to compound 17 in a coupling reaction with compound 10 using a coupling reaction agent, for example TPTU / HOBT or EDC, in the presence of an inert solvent, for example dichloromethane, and a tertiary amine, for example diisopropylethylamine, in order to obtain a coupling product Compound 17.

Compound 17 is converted to compound I by deprotection in a strongly acidic environment, for example in hydrochloric acid, in the presence of a polar organic solvent, for example dioxane, so as to obtain compound I without protection.

The compound of the present invention is a base, such a compound is suitable for use in the form of a free base or in the form of a pharmaceutically acceptable acid addition salt thereof.

Acid addition salts may be a more convenient form for use, and in practice, the use of the salt form essentially means the use of the free base form. Acids that can be used to prepare acid addition salts include preferably those which, when mixed with the free base, result in pharmaceutically acceptable salts, that is, salts whose anions are non-toxic to the patient in pharmacological doses of salts, so that the beneficial effects of inhibition, inherent in the free base, are not distorted by the side effects attributed to anions. Although pharmaceutically acceptable salts of these basic compounds are preferred, all acid addition salts are useful sources of the free base form, even if a particular salt, by itself, is only needed as an intermediate, for example, if the salt is obtained solely for purification and identification or if it is used as an intermediate in the preparation of a pharmaceutically acceptable target by ion exchange processes. Pharmaceutically acceptable salts that are within the scope of the present invention include those derived from mineral and organic acids, and include hydrohalides, for example hydrochloride and hydrobromide, sulfates, phosphates, nitrates, sulfamates, acetates, citrates, lactates, tartrates, malonates, oxalates, salicylates, propionates, succinates, fumarates, maleates, methylene bis-beta-hydroxy naphthoates, benzoates, tosylates, gentisinates, isethionates, di-p-toluoyl tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates xylsulfamates and hinates. A more specific example of a salt of a compound of the formula is a hydrochloride salt.

In addition to being able to use the salts of the compounds of the invention themselves as active compounds, they are also suitable for the purification of the compound, for example, by using the difference in solubility between the salts and the starting compound, by-products and / or starting material using techniques known to those skilled in the art.

According to yet another aspect of the invention, acid addition salts of a compound of the present invention can be prepared by reacting the free base with a suitable acid using or adapting known methods. For example, salts of a compound of the present invention formed by the addition of acids can be prepared either by dissolving the free base in water or an aqueous alcoholic solution, or in other suitable solvents containing the appropriate acid, and isolating the salt by evaporating the solution, or by reacting the free base and acid in organic solvent, in this case, the salt is separated directly or can be obtained by concentration of the solution.

The acid addition salts of a compound of the present invention can be recovered from the salts using or adapting known methods. For example, the starting compound of the present invention can be obtained from its salts formed by the addition of acids, treatment with alkalis, for example aqueous sodium bicarbonate or aqueous ammonia.

Starting materials and intermediates can be obtained using or adapting known methods, for example, the methods described in the reference examples, or their obvious chemical equivalents.

The present invention also relates to some of the intermediates from the above schemes, and the processes for their preparation described herein are themselves additional features of the present invention.

Examples

For a better understanding of the present invention, reference may be made to the following preferred, but not the only examples, which are given as an illustration of the invention. The following examples are provided to more fully illustrate preferred embodiments of the invention. However, it should in no way be construed as limiting the broad scope of the present invention.

In the spectra of nuclear magnetic resonance (NMR) below, the values of chemical shift are expressed in ppm. relative to tetramethylsilane. The abbreviations have the following meanings: w = wide, dd = doublet of doublets, s = singlet; m = multiplet.

EXAMPLE 1

Stage 1: obtaining 1- (3-bromo-4-fluorobenzyl) -2,2,5,5-tetramethyl [1,2,5] azadisilolidine (compound 3)

110 g (0.46 mol) of 3-bromo-4-fluorobenzylamine hydrochloride (2) is suspended in 900 ml of methylene chloride in a 2-liter three-necked round bottom flask equipped with a nitrogen jacket, a thermocouple temperature sensor with a Teflon coating and a mechanical stirrer, and cooled to ~ 5 ° C in an ice bath. A total of 144 g (1.42 mol, 3.1 equiv. D = 0.7) is added, resulting in a thick suspension. Then, a solution of 1,2-bis (chlorodimethylsilyl) ethane (100 g, 0.46 mol) in 250 ml of methylene chloride is poured dropwise for more than 1.5 hours, while maintaining the temperature of the reaction mixture between 5 and 8 ° C. The mixture was stirred for 30 minutes and then warmed to room temperature. The precipitate of triethylamine hydrochloride is filtered off. The filtrate was concentrated in vacuo (40 ° C, <50 mbar), 1 L of pentane was added and an additional precipitate of Et ₃ N × HCl was collected by filtration, which formed. The process is repeated once more with 1 l of pentane. The filtrate was concentrated in vacuo (40 ° C, <5 mbar) to a colorless liquid, which crystallized upon cooling to form 159 g (~ 100%) of compound 3 as a white crystalline powder.

The completion of the reaction must be monitored by ¹ H NMR to ensure that there are virtually no residues of starting material 2 and triethylamine hydrochloride, since they lead to the consumption of n-butyl lithium in the next step, thereby reducing the yield of the reaction.

Step 2: Preparation of 1-benzyl-4- [2-fluoro-5- (2,2,5,5-tetramethyl [1,2,5] azadisylolidin-1-ylmethyl) phenyl] piperidin-4-ol (compound 5 )

A solution of compound 3 (159 g, 0.4574 mol) in 1.5 L of anhydrous THF was placed in a 5-liter three-necked round bottom flask equipped with a nitrogen jacket, a thermocouple temperature sensor with a Teflon coating and a mechanical stirrer, and cooled to -75 ° C. A 2.5 M solution of n-butyllithium (192 ml, 0.48 mol, 1.05 equiv.) Was added dropwise to this stirred solution over about 1 hour, while maintaining the temperature of the reaction mixture between -72 and -75 ° C. After 30 minutes, a solution of 1-benzyl-4-piperidone (compound 4; 88.2 g, 0.47 mol, 1.02 equiv.) In anhydrous THF (350 ml + 50 ml for washing) was added dropwise over more than 1 hours, while maintaining the reaction temperature below -70 ° C. After stirring for 20 min at a temperature of from -70 to -75 ° C, 200 ml of methanol are added to the reaction mixture (color changes from orange to yellow), then the reaction mixture is allowed to warm to room temperature. The reaction mixture was concentrated in vacuo (50 ° C) to obtain 295 g of a brown oily liquid, compound 5.

Stage 3: obtaining 4- (5-aminomethyl-2-fluorophenyl) -1-benzylpiperidin-4-ol (phosphate salt) (compound 6)

The crude compound 5 (~ 295 g) was diluted with 1 L of methylene chloride and transferred to a 3-liter three-necked round bottom flask equipped with a Teflon-coated thermocouple temperature sensor and a mechanical stirrer. 53 g of 85% H ₃ PO ₄ (1 equiv.) Were slowly added to this solution with stirring (exothermic reaction) and the mixture was concentrated in vacuo (40 ° C). 1 L of methylene chloride was added and the mixture was concentrated in vacuo (40 ° C, 1 mbar) to a yellow foam, which was then stirred in 1 L of heptane. A precipitate forms, which is separated by filtration. The brown precipitate was dried to give 190 g of compound 6. MS: m / z 315 (M + H) was detected.

Stage 4: obtaining 3- (1-benzyl-1,2,3,6-tetrahydropyridin-4-yl) -4-fluorobenzylamine (compound 7)

All 190 g of dried Compound 6 was transferred to a 2-liter, three-necked round bottom flask equipped with a thermocouple temperature sensor with a Teflon coating and a mechanical stirrer. A total of 600 ml of 85% H ₃ PO _{4 is} added. The resulting suspension is gradually heated to 100 ° C with stirring. The completion of the reaction is monitored by HPLC (usually 2 to 3 hours). After completion of the reaction, the reaction mixture was cooled to room temperature and diluted with 800 ml of water. The aqueous layer was washed with ether (2 × 200 ml). The aqueous layer is then neutralized with 50% aq. NaOH to pH> 9, while maintaining the temperature below 30 ° C. Such an aqueous solution is a highly concentrated buffer system, the pH of which remains unchanged for about 7-8, until the point of neutralization is reached. Confirmation of the completion of neutralization is obtained by adding a few drops of the base to a small sample of the supernatant to ensure that precipitation stops. A significant amount of salt precipitates. The mixture was filtered and the precipitate was washed with DCM (dichloromethane) (about 3 L) and 2 L of water. The organic layer (lower) is separated and washed with water (2 × 1 L). The organic solution was concentrated in vacuo (40 ° C) to give 107 g of a brown oily liquid, compound 7. MS: m / z 297 (M + H) was detected.

Step 5: Preparation of [3- (1-benzyl-1,2,3,6-tetrahydropyridin-4-yl) -4-fluorobenzyl] carbamic acid tert-butyl ester (compound 8)

50 g of compound 7 and a solution of methanol (800 ml) and water (400 ml) are placed in a 2-liter three-necked round-bottom flask equipped with a thermocouple temperature sensor with a Teflon coating and a mechanical stirrer. 36.8 g of BOC anhydride and 2 ml of 50% NaOH are added and the mixture is stirred at room temperature. The product precipitates out of solution after 2 hours of stirring. If the reaction product with BOC does not precipitate, the upper aqueous layer is decanted and n-heptane is added to the oily liquid to crystallize the product. The resulting mixture was stirred overnight at room temperature. The precipitate was filtered off and stirred in 1.05 L MeOH / water (2: 1 by volume) for 4 hours, then filtered off and dried for 4 days to obtain 40 g (total yield is about 35 to 48% in compound 3) of compound 8 in the form of a light yellow powder. Purity 96.3% by HPLC. MS: m / z 397 (M + H), 398 (M + 2H).

Stage 6: obtaining the acetate salt of tert-butyl ether 6- (4-fluoro-3-piperidin-4-ylbenzyl) carbamic acid (compound 9)

64 g (0.16 mol) of compound 8, 6.4 g of palladium hydroxide / C 20%, 19.5 g (2 equiv.) Of glacial acetic acid and 250 ml of methanol are loaded into a 1 liter hydrogenation vessel. The reaction vessel was purged (N ₂ / vacuum, 3 times), then filled with H ₂ to 40 psig. inch and shake overnight at room temperature. The catalyst was filtered off and the filtrate was concentrated in vacuo (40 ° C) until an oily liquid was obtained. 200 ml of isopropyl ether was added and stirred overnight. The white precipitate, which precipitates in this case, is separated by filtration, washed with isopropyl ether and dried, so as to obtain 53.5 g of a white powder with an HPLC purity of 95.4%. The powder was stirred in 500 ml of MTBE for 5 hours, then separated by filtration and again stirred in 500 ml of MTBE overnight. The powder was separated by filtration and dried to obtain 51.3 g (86.3%) of the white acetate powder of compound 9. Elemental analysis: Calculation for C ₁₇ H ₂₅ FN ₂ O ₂ : C 61.94; H 7.93; N, 7.6. Found: C, 62.0; H 8.17; N, 7.49. According to Karl Fischer: 0.34% of water. HPLC: Rt 9.14 min, 97% purity by area under the curve. MS: m / z 309 (M + H), 310 (M + 2H).

Step 7: Preparation of (4-Fluoro-3-piperidin-4-ylbenzyl) carbamic Acid Tert-Butyl Ether (Compound 10)

The acetate of compound 9 (51 g) was dissolved in 400 ml of water and the pH was adjusted to 5 by adding 2N HCl. The aqueous solution was washed with ether (2 × 200 ml). The aqueous layer is neutralized to a pH> 12 to 50% aq. NaOH and extracted with ether (2 × 300 ml). The organic layer was washed with water and dried over Na ₂ SO ₄ , and then concentrated to an oily liquid. 200 ml of n-pentane are added to an oily liquid and stirred for 3 hours. The solid product was separated by filtration, washed with n-pentane and dried at room temperature under standard vacuum for 24 hours to obtain 42 g of compound 10 (98%) as a white powder. Electrospray ionization mass spectroscopy, m / z 309 (M + H). Elemental analysis: Calcd for C ₁₇ H ₂₅ FN ₂ O ₂ : C 66.21; H 8.17; N, 9.08. Found, not adjusted for water: C 64.30; H 8.64; N, 8.77. According to Karl Fischer: 2.57% of water. HPLC: Rt 9.16 min, 98.1% purity by area under the curve.

Note. If the purity of the acetate of compound 9 by HPLC is less than 95%, instead of pure n-pentane, a mixture of n-pentane and ether (up to 1: 1 by volume) can be used to crystallize the product (free base) from an oily liquid. The product is very soluble in ether, so if more ether is used, even more product will be lost, thereby reducing yield.

EXAMPLE 2

Stage 1: obtaining bis (propoxythiocarbonyl) sulfide (compound 11)

84.2 g of potassium hydroxide powder (1.27 mol) was added to 530 ml of n-propanol, placed in a three-necked round bottom flask equipped with a mechanical stirrer and a cooling bath, at room temperature. Then, 80 ml of carbon disulfide (1.33 mol) are added dropwise to the solution over a period of more than 1 hour through an addition funnel with equalization of pressure. Stirring is continued for 3 hours. 200 ml of water are added. 73.5 g (0.6 mol) of pure propyl chloroformate are poured dropwise through an addition funnel with pressure equalization. The resulting mixture was stirred overnight at room temperature. The mixture is then diluted with 400 ml of heptane. The aqueous layer was extracted twice with 100 ml of heptane. The combined organic layer was washed with 100 ml of water, 100 ml of brine, and then dried over potassium carbonate and evaporated. Residual propanol is removed by distillation under high vacuum to obtain a clear yellow liquid. 10 g of a clear yellow liquid was dissolved in 10 g of heptane and purified by silica gel chromatography eluting with heptane to obtain 8.35 g of compound 11 (41% yield).

Stage 2: Obtaining O-propyl ester of 3-oxoethiobutyric acid (compound 12)

1.82 g (45 mmol) of sodium hydride are suspended in 40 ml of toluene. To the suspension obtained, a solution of 1.74 g (30 mmol) of acetone and 4.76 g (20 mmol) of compound 11 in 10 ml of toluene at 40 ° C is poured with stirring. The reaction is initiated by introducing a small amount of potassium hydride. In this case, bubbles are released, and the color of the reaction mixture changes from yellow to orange. The reaction mixture was stirred at 40 ° C for one hour, and then cooled to 0 ° C in an ice water bath. The reaction mixture was then poured into a beaker containing 11 ml of 4N HCl, ice and 150 ml of ether. The organic layer was separated and concentrated. The crude product was purified by silica gel chromatography, eluting with 5% ethyl acetate in heptane to give 3-oxo-thiobutyric acid O-propyl ester (compound 12).

Step 3: Preparation of (3-oxo-1-propoxybut-1-enylsulfanyl) acetic acid methyl ester (compound 13)

Compound 12 is then dissolved in 40 ml of DMF and cooled to 0 ° C. 4.6 g of alpha-bromomethyl acetate in 5 ml of DMF and 5.2 ml of triethylamine are added to the resulting solution. Instant formation of a white precipitate occurs. The resulting suspension was stirred at 0 ° C for 2 hours, and then poured into ether / ice-water. The organic layer was separated and concentrated. The crude product was purified by silica gel chromatography and then eluted with 3% methanol in DCM to give 2.61 g of compound 13 (56% yield in two steps).

Stage 4: obtaining methyl ester of 3-methyl-5-propoxythiophene-2-carboxylic acid (compound 14)

A mixture of 2.61 g (11.24 mmol) of compound 13 and 2 ml of a 0.5 M sodium methylate / methanol mixture in 40 ml of methanol is heated at 70-73 ° C for 40 minutes. The mixture was then poured into ether / ice-water. The organic layer was separated and concentrated. The crude product was purified by silica gel chromatography eluting with DCM to give 1.85 g (76.8% yield) of compound 14.

Step 5: Preparation of 4-bromo-3-methyl-5-propoxythiophene-2-carboxylic acid methyl ester (compound 15)

To a solution of 1.22 g (5.7 mmol) of compound 14 in 20 ml of trichloromethane is added 11 ml of a 0.55 M mixture of bromine / trichloromethane at 0 ° C and stirred for 10 minutes. The mixture was quenched with an aqueous solution of sodium sulfite and then extracted with DCM. The organic layer was separated and concentrated. The crude product is purified by passing through a layer of potassium carbonate-silica gel and washed with DCM to give 1.67 g (100% yield) of compound 15.

Stage 6: obtaining 4-bromo-3-methyl-5-propoxythiophene-2-carboxylic acid (compound 16)

To a solution of 1.67 g of compound 15 (5.70 mmol) in 27 ml of dioxane was added 10 ml of 2 M LiOH / water and 9 ml of water. The resulting mixture was stirred for 4 hours at room temperature. The mixture is then diluted with 10 ml of water and extracted with 10 ml of ether twice. The aqueous solution was cooled in an ice water bath and acidified with 4 M HCl. White powder was collected by suction filtration to obtain 1.28 g (80.5% yield) of compound 16.

EXAMPLE 3

Step 1: Preparation of {3- [1- (4-bromo-3-methyl-5-propoxythiophen-2-carbonyl) piperidin-4-yl] -4-fluorobenzyl} carbamic acid tert-butyl ester (compound 17)

To a solution of 170 mg of compound 16 (0.61 mmol) in 25 ml of DCM in a stream of N ₂ add 170 mg of TPTU (0.61 mmol) and 80 mg of 1-hydroxy-1H-benzotriazole (HOBt) (0.61 mmol). Stirring is continued for 3 minutes. A solution of 200 mg (0.65 mol) of compound 10 in 5 ml of DCM and 0.3 ml of diisopropylethylamine (1.2 mmol) is then poured onto the mixture. The resulting mixture was stirred for 24 hours at room temperature. The mixture was then washed with 20 ml of water, dried over anhydrous sodium sulfate and concentrated. The crude oily product was purified by silica gel chromatography eluting with 3% methanol in DCM to give 0.3 g (86.5% yield) of compound 17.

¹ H NMR (CDCl _3, TMS): δ 7.14-7.04 (m, 2H), 6.95 (dd, H), 4.82 (br.s, H), 4.38 (br. d, 2H), 4.23 (d, 2H), 4.05 (t, 2H), 3.17-2.95 (m, 3H), 2.21 (s, 3H), 1.92-1 59 (m, 6H); 1.44 (s, 9H); 1.05 (t, 3H). MS (ESI +): 569 (M ⁺ +1).

Step 2: Preparation of [4- (5-Aminomethyl-2-fluorophenyl) piperidin-1-yl] - (4-bromo-3-methyl-5-propoxythiophen-2-yl) methanone hydrochloride (compound I)

A solution of 0.25 g of compound 17 (0.44 mmol) in 8 ml of 4 M HCl / dioxane was stirred under a stream of nitrogen at room temperature for 3 hours. The solution was diluted with 30 ml of ether and stirred for 5 minutes. The liquid above the precipitate is decanted. The precipitate is washed with 30 ml of ether and the liquid is again decanted. Then the precipitate was dissolved in 3% methanol in DCM and purified by silica gel chromatography, eluting with 3 to 10% methanol in DCM. Compound I, 0.19 g (yield 86.4%), was obtained as an amorphous substance, removing the solvent from the combined pure fractions of the product by evaporation. The product is an amorphous glass.

¹ H NMR (CDCl _3, TMS): δ 8,62 (. Br s, 3H), 7,53-7,43 (m, H), 7,36-7,26 (m, H), 6 97 (dd, H), 4.22 (brd, 2H), 4.18-3.99 (m, 4H), 3.16-2.90 (m, 3H), 2.17 (s, 3H), 2.00-1.60 (m, 6H), 1.02 (t, 3H). MS (ESI +): 469 (M ⁺ +1). 100% purity by LC / MS (UV 220 nm and total ion count).

BIOLOGICAL ACTIVITY

The properties of the compounds of the present invention are also demonstrated by the examples: 1) its inhibitory ability against beta-tryptase (IC ₅₀ and K _i values) and 2) its activity measured in a model of airway hyperresponsiveness in guinea pigs (oral average effective dose ED ₅₀ ) .

TEST PROCEDURE “IN VITRO”

Since all the effects of tryptase, as described in the section on the background of the invention, depend on its catalytic activity, compounds that inhibit its catalytic activity will potentially inhibit the effect of tryptase. The inhibition of such catalytic activity can be measured using enzyme analysis and in vitro cell analysis.

Inhibition of tryptase activity is confirmed by the use of either tryptase isolated from human lungs or recombinant human beta-tryptase expressed in yeast cells. Essentially equivalent results are obtained using an isolated native enzyme or an expressed enzyme. The assay used a 96-well microplate (Costar 3590) with L-pyroglutamyl-L-prolyl-L-arginine-p-nitroanilide (S2366: Quadratech) as a substrate (mainly as described in McEuen et. Al. Biochem Pharm, 1996, 52, pp. 331-340). The analysis is performed at room temperature using 0.5 mM substrate (2 × Km), and these microplates are obtained using a microplate reader (Beckman Biomek Plate reader) at a wavelength of 405 nm.

Materials and methods for primary tryptase screening (chromogenic analysis)

Analytical buffer

50 mM Tris (pH 8.2), 100 mM NaCl, 0.05% Tween 20, 50 μg / ml heparin.

Substrate

S2366 (stock solutions 2.5 mM).

Enzyme

Stock solutions of purified recombinant beta-tryptase 310 μg / ml.

Protocol (Single Point Definition)

Add 60 μl of the diluted substrate (final concentration of 500 μM in assay buffer) to each well.

Add the compound (two cells each), final concentration 20 μM, volume 20 μl.

Add the enzyme at a final concentration of 50 ng / ml, volume 20 μl.

The total volume for each cell is 100 μl.

Shake for a short time and incubate at room temperature in the dark for 30 minutes.

Read absorbance at 405 nm.

The following control samples are provided for each of the cells:

total: 60 μl of substrate, 20 μl of buffer (with a final concentration of 0.2% DMSO), 20 μl of enzyme,

non-specific: 60 μl of substrate, 40 μl of buffer (with 0.2% DMSO),

total: 60 μl of substrate, 20 μl of buffer (without DMSO), 20 μl of enzyme,

nonspecific: 60 μl of substrate, 40 μl of buffer (without DMSO).

Protocol (IC definition _fifty  and K _i )

The protocol is essentially the same as described above, except that the compound is added in two cells with the following final concentrations: 0.01, 0.03, 0.1, 0.3, 1, 3, 10 μM (all dilutions are made manually). For each assay, as for a point, and for determining the IC _50, the reference compound used to obtain IC ₅₀ for comparison. From the value of IC ₅₀ , K _i can be calculated using the following formula: K _i = IC ₅₀ / (1 + [substrate] / Km).

The inhibitory ability of beta-tryptase for the compounds of formula I is determined by the values of IC ₅₀ and K _i for 76 nm and 15 nm, respectively.

TEST PROCEDURE “IN VIVO”

Analysis protocol

Sensitization and use of the drug. Hartley male guinea pigs (225-250 g) are sensitized with egg albumin (0.5 ml of a 1% solution, intraperitoneally and subcutaneously). On day 4, the animals received a repeated dose (intraperitoneally) of 0.5 ml of 1% egg albumin. On day 21, animals received an oral dose (2 ml / kg) of either a vehicle (0.5% methyl cellulose / 0.2% Tween 80) or a test compound 2 hours before antigenic stimulation. Thirty minutes before antigenic stimulation, mepiramine (30 mg / kg, ip) was also administered to animals to prevent anaphylactic collapse. Then the animals were subjected to aerosol saline solution (control animals) or 1% egg albumin for 5 minutes using the deVilbiss Ultraneb aerosol apparatus.

Assessment of airway hypersensitivity: within eighteen to twenty-four hours after stimulation, the animals were anesthetized using a combination of ketamine (133 mg / kg) and xylazine (24 mg / kg) administered intramuscularly, surgical preparation was performed and recorded in a general plethysmograph to determine lung function . The animals were connected to Ugo-Basile mechanical ventilators providing a tidal volume of 1 ml / 100 g at a frequency of 50 breaths / minute through a tracheal cannula. A cannula for histamine stimulation was also introduced into the jugular vein. An esophageal cannula filled with water was placed in such a way as to allow the detection of transpulmonary pressure. Transpulmonary pressure is determined by the difference between the tracheal and esophageal cannulas using a differential pressure sensor. Volume, air flow, and transpulmonary pressure were monitored using a pulmonary analysis system (Buxco XA software) and used to calculate pulmonary resistance (cm H ₂ O / ml · sec) and dynamic extensibility (ml / cm H ₂ O). Airway resistance and dynamic extensibility are calculated for each breath. Histamine is administered intravenously, and reactivity with respect to increasing concentrations (0.3–20 μg / kg) is calculated. ED _50s are estimated by the area under the curve obtained for individual histamine dose-response curves.

Plasma and lung levels of the drug

In satellite groups, plasma and lung concentration levels of the compound were determined. Three to four guinea pigs from each treatment group of the experimental drug were used to determine drug levels. At a given point in time (2 or 24 hours after administration), animals were killed and 1 ml of a blood sample was taken by heart puncture, collecting them in heparin-coated syringes containing 20 μl (per 1 ml of blood) 5 mM hydralazine solution. Plasma was separated from the cellular component of the blood by centrifugation and stored at -20 ° C until analysis. Lung samples are prepared with release from connective tissues, blotted dry, weighed and stored in 20 ml vials containing 5 ml of a 5 mM hydralazine solution in physiological saline. Samples of frozen plasma are then transferred onto dry ice to samples of the experimental pharmacokinetics group to determine compound concentration levels.

results

The effectiveness of compound I is determined by the hypersensitivity of the respiratory tract to histamine in sensitized guinea pigs, by oral route. Compound I has no effect on basal airway resistance or basal dynamic lung extensibility.

Sensitization and single stimulation with egg albumin leads to an increase in bronchial reactivity to histamine, which is recorded as a left shift in the spasmogen dose-response curve and as a noticeable increase in the area under the curve for airway resistance and lung distensibility. Absolute values reflect an increase in airway resistance and a decrease in lung distensibility.

When administered orally 2 hours before stimulation with egg albumin, compound I provides significant protection against antigen-stimulated hypersensitivity of the upper respiratory tract with ED ₅₀ = 0.1 mg / kg, as shown by estimates of airway resistance and dynamic lung distensibility.

In a separate group of animals, the levels of compound I are determined in bronchoalveolar lavage (BAL), lungs and plasma 2 hours after administration of the compound (1 mg / kg, oral). A noticeable amount of compound I is recorded in target organs, for example, in the lungs and BAL (without taking into account the dilution factor for BAL). In plasma, much lower levels of compound are detected.

In the satellite group of animals 24 hours after administration, the levels of compound I were also measured in both lungs and plasma. While it was not possible to detect Compound I in the plasma, noticeable dose-dependent significant amounts were recorded in the lungs of guinea pigs 24 hours after administration. It was also shown that compound I has a long-lasting effect with an average ED _{50 of} 0.4 mg / kg when administered 24 hours before antigen stimulation. This duration of action is in agreement with its extended long-term exposure.

Oral administration data of a compound of the present invention in a guinea pig hypersensitivity model of airways clearly demonstrate that the compound exhibits tryptase inhibitory activity. Therefore, the compound of the present invention easily finds use as a pharmaceutical preparation for the treatment of a wide range of diseases associated with tryptase, and naturally in methods of treating such diseases in a patient.

The present invention should not be limited in scope to those specific implementations that are described herein. In fact, various modifications of the invention, in addition to those already described in the present description, will be apparent to those skilled in the art from the above description and the attached drawings. Such modifications are intended to fall within the scope of the appended claims.

In the present description, various publications are cited, which, by reference to them, are fully included in the presented text.

Claims (13)

1. The compound of formula I

or a pharmaceutically acceptable salt of said compound. 2. The compound according to claim 1 in the form of a pharmaceutically acceptable salt thereof. 3. The compound of claim 2, wherein the pharmaceutically acceptable salt is hydrochloride. 4. A method of treating patients suffering from or susceptible to physiological diseases in which an increase in the level of a tryptase inhibitor is necessary, comprising administering to the patient a therapeutically effective amount of a compound according to claim 1. 5. The method according to claim 4, in which the physiological disease is selected from the group including inflammatory disease, ophthalmic conjunctivitis, spring conjunctivitis, irritable bowel syndrome, asthma, allergic rhinitis, fibrosis, scleroderma, pulmonary fibrosis, cirrhosis, myocardial fibrosis, neurofibroma , hypertrophic scars, dermatological disease, disease associated with rupture of atherosclerotic plaques, periodontal disease, diabetic retinopathy, tumor growth, anaphylaxis and multiple sclerosis. 6. The method according to claim 5, where the physiological disease is an inflammatory disease. 7. The method according to claim 6, where the inflammatory disease is arthritis. 8. The method according to claim 5, where the physiological disease is a dermatological disease. 9. The method of claim 8, where the dermatological disease is atopic dermatitis or psoriasis. 10. The method according to claim 5, where the physiological disease is associated with rupture of atherosclerotic plaques. 11. The method according to claim 10, where the rupture of atherosclerotic plaques leads to myocardial infarction, stroke or angina pectoris. 12. The method according to claim 4, whereby the introduction is carried out in such a way that the compound according to claim 1 is preferably distributed in the tissues of the lungs, and not in the plasma. 13. A pharmaceutical composition having an inhibitory effect on beta-tryptase, comprising a therapeutically effective amount of a compound according to claim 1 and a pharmaceutically acceptable excipient thereof.

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